Nov 22, 1993 - DNA to which GJB1 and CCG1 have already been mapped. A recent report of mutations in the GJB1 gene in subjects with CMTX1 makes this a ...
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X linked Charcot-Marie-Tooth disease (CMTX1): a study of 15 families with 12 highly informative polymorphisms S Cochrane, J Bergoffen, N D Fairweather, E Muller, M L Mostacciuolo, A P Monaco, K H Fischbeck, N E Haites
males showed pes cavus, distal muscle wasting, and weakness which is progressive from the mid teens. Diminished or absent tendon reflexes are also present, initially in the lower limbs but eventually progressing to the upper limbs, with variable sensory loss. In addition, affected males within these families had median nerve conduction velocities of less than 38 m/sec, a feature also observed in dominantly inherited CMT type 1. Wide variation in the severity of clinical signs was found in carrier females who had later onset of symptoms, with some obligate carriers being asymptomatic. Genomic DNA was isolated from peripheral white blood cells using a modified standard technique."3 Southern blot analysis was performed using the markers M27P (DXS225)"4 and PGK1. 5 PCR amplification of microsatel(J7 Med Genet 1994;31:193-196) lites at HAR,'6 PGKP1,'7 DXS106,'8 DXS135,19 DXS453,20 DXS559,2' DXS227,'5 DXS56 and PGK1,22 DXS441,23 and X linked Charcot-Marie-Tooth disease DXYS 124 was performed according to the con(CMTX) is a clinically heterogeneous group of ditions previously described. diseases with both dominant and recessive Affected members of all families were also inheritance. The dominant form (CMTX1) probed with pVAW409R3a,"' to confirm the was provisionally assigned to the proximal absence of the chromosome 17pI 1.2-12 duplilong arm of the X chromosome in 1985.1 This cation. has been confirmed by the work of several other groups in many families,2-8 and the gene has subsequently been localised to the segment Results and discussion of proximal Xq between PGKP1 (Xql 1.2-12) All the families show a pattern of inheritance and DXS72 (Xq21.1).9 In addition, other consistent with an X linked disease (that is, no workers have described recessive forms, one male to male transmission). DNA analysis of associated with mental retardation, provision- affected members no indication of the ally mapping to Xp22.2 and the other to duplication at 17p1showed 1.2-12, increasing the likeXq26.'0 lihood X of linked inheritance. CMTX1 was once considered rare in comNot all of the polymorphic markers studied parison to autosomal dominant forms. How- were informative in every family. As expected, ever the ability to test for the duplication at there were fewer recombinants detected in the chromosome 17pl 1.2-12,"1 12 characteristic of smaller pedigrees. Clinically normal females the majority of HMSN1A families, has who were carriers were excluded allowed diagnosis to be reconsidered in many from the not obligate The individual recombinaanalysis. smaller families, whose inheritance and clinical tion events which did occur in specific families features were not inconsistent with either are shown in 4. These recombinants localise fig mode of inheritance. the X linked CMT to the region distal to gene We report here a study performed on 15 X DXS106 and proximal to (Xqll.2-12)'926 linked dominant families (figs 1A, 1B, and 2) (Xql3.1).25 These markers flank apusing 12 recently described highly informative DXS559 2 to 3 Mb of DNA (A P Monaco, proximately polymorphisms (fig 3) spanning the region of personal communication, 1993). interest on the X chromosome. For diagnostic purposes, the locus DXS453 would therefore appear to be a useful marker for the disease, with no recombinations at this Materials and methods site having yet been detected. The families were identified at regional neuroTwo genes have already been mapped to the logical and genetic clinics in the United King- region Xql3.1, GJB127 and CCG1.2628 The dom, the USA, and Italy. Typically, affected CCG1 (cell cycle Gl phase defect) gene comAbstract X linked dominant Charcot-Marie-Tooth disease (CMTX1) has previously been localised to Xql3-21. Fifteen -families were studied using 12 highly informative polymorphisms in the pericentric region of the X chromosome. Phase known recombinations in these families localise the X linked dominant CMT gene to the region distal to DXS106 (Xqll.2-12) and proximal to DXS559 (Xql3.1). These markers flank approximately 2 to 3 Mb of DNA to which GJB1 and CCG1 have already been mapped. A recent report of mutations in the GJB1 gene in subjects with CMTX1 makes this a strong candidate gene.
Medical Genetics, Department of Molecular and Cell Biology, University of Aberdeen Medical School, Foresterhill, Aberdeen AB9 2ZD, UK S Cochrane N D Fairweather N E Haites
Department of Genetics, Kaiser Permanente Medical Group, San Jose, CA, USA J Bergoffen Department of Neurology, University of Pennsylvania School of Medicine, Philadelphia, PA, USA J Bergoffen K H Fischbeck ICRF Laboratories, Institute of Molecular Medicine, John Radcliffe Hospital, University of Oxford, Oxford, UK N D Fairweather A P Monaco Genetics Laboratory, Department of Biology, University of Padova, Padova, Italy E Muller M L Mostacciuolo Correspondence
to
Dr Haites. Received 6 October 1993 Revised version accepted for publication 22 November 1993
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Cochrane, Bergoffen, Fairweather, Muller, Mostacciuolo, Monaco, Fischbeck, Haites A
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(B) Pedigrees of eight British and one Italian families.
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X linked Charcot-Marie- Tooth disease (CMTXI)
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Figure 3 Schematic map of the X chromosomal region Xp21.1-Xq2I.3, showing loci segregated into 15 different intervals," with relevant marker positions.
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Figure 4 Recombination analysis using variable number simple sequence repeats (VSSR) and restriction fragment length polymorphisms (RFLP). The markers used are listed in chromosomal order25 on the left. Across the top, the numbers represent carrier females with informative meioses. The vertical lines represent X chromosomes that have undergone phase known recombination during meiosis. Open circles represent non-recombinant loci, and closed circles represent recombinations that have occurred.9 A horizontal bar denotes an uninformative result. The arrows point in the direction in which the X linked HMSN gene must therefore lie.
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plements a hamster cell cycle mutation and is therefore required in normal cell growth and division. Its functional presence allows cells to pass out of the Gl phase of cell cycle events, which in turn commits them to completing the S, G2, and M phases.29 GJB1 (gap junction protein f1), otherwise known as Connexin32 (Cx32), is, as its name suggests, a gap junction protein of molecular weight 32 kDa. Cx32 is widely expressed in human tissues30 and mutations within this gene have been found in subjects with CMTX1 making this a strong candidate gene for this disease.3' This work was funded by grant GENO-CT91-0017 from the Commission of the European Communities and previously by the Muscular Dystrophy Group of Great Britain and Northern Ireland, the Muscular Dystrophy Association, the Dana Foundation, and by NINDS grants NS08075 and NS26330. The financial support of Teleton-Italy is also gratefully acknowledged. We would like to thank all of the doctors and families who have contributed to this study. 1
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Gal A, Mucke J, Theile H, et al. X linked dominant Charcot-Marie-Tooth disease: suggestions of linkage with a cloned DNA sequence from the proximal Xq. Hum Genet 1985;70:38-42. Beckett J, Holden JJA, Simpson NE, et al. Localization of X linked dominant Charcot-Marie-Tooth disease (CMT2) to Xql3. J7 Neurogenet 1986;3:225-31. Fischbeck KH, Ar-Rushdi N, Pericak-Vance M, et al. X linked neuropathy: gene localization with DNA probes. Ann Neurol 1986;20:527-32. Rozear MP, Pericak-Vance MA, Fischbeck K, et al. Hereditary motor and sensory neuropathy, X linked: a half century follow-up. Neurology 1987;37:1460-5. Ionasescu VV, Burns TL, Searby C, Ionasescu R. X linked dominant Charcot-Marie-Tooth neuropathy with 15 cases in a family genetic linkage study. Muscle Nerve 1988;1 1:1 154-6. Goonewardena P, Wellhinda J, Anvret M, et al. A linkage study of the locus for X linked Charcot-Marie-Tooth disease. Clin Genet 1988;33:435-40. Haites N, Fairweather N, Clark C, et al. Linkage in a family with X linked Charcot-Marie-Tooth disease. Clin Genet 1989;35:399-403. Mostacciuolo ML, Muller E, Fardin P, et al. X linked Charcot-Marie-Tooth disease. A linkage study in a large family by using 12 probes of the pericentromeric region. Hum Genet 1991;87:23-7. Bergoffen J, Trofatter J, Pericak-Vance MA, et al. Linkage localization of X linked Charcot-Marie-Tooth disease. Am Hum Genet 1993;52:312-18. Ionasescu VV, Trofatter J, Haines JL, et al. Heterogeneity in X linked recessive Charcot-Marie-Tooth neuropathy. AmJ Hum Genet 1991;48:1075-83. Racymaekers P, Timmerman V, Nelis E, et al. Duplication
in chromosome 17p1 .2 in Charcot-Marie-Tooth neuropathy type la(CMTla). Neuromusc Dis 1991;1:93-7. 12 Lupski JR, Montes de Oca-Luna R, Slaugenhaupt S, et al. DNA duplication associated with Charcot-Marie-Tooth disease type IA. Cell 1991;66:219-32. 13 Kunkel LM, Smith KD, Boyer SH, et al. Analysis of human Y-chromosome specific reiterated DNA in chromosome variants. Proc Natl Acad Sci USA 1977;74: 1245-9. 14 Fraser NJ, Boyd Y, Brownlee GG, Craig IW. Multi-allelic RFLP for M27f, an anonymous single copy genomic clone at Xpl 1.3-Xcen. Nucleic Acids Res 1987;15:9616. 15 Hutz MH, Michelson AM, Antonarakis SE, et al. Restriction site polymorphism in the phosphoglycerate kinase on the X chromosome. Hum Genet 1984;66:217-19. 16 La Spada AR, Wilson EM, Lubahn DB, et al. Androgen receptor gene mutations in X linked spinal and bulbar muscular atrophy. Nature 1991;352:77-9. 17 Browne DL, Zonana J, Litt M. Dinucleotide repeat polymorphism at the PGK1P1 locus. Nucleic Acids Res 1992;20:1 169. 18 Fairweather N, Chelly J, Monaco AM. Dinucleotide polymorphisms from DXS106 and DXS27 YACs using a two stage approach. Hum Mol Genet 1993;2:607-8. 19 Markiewicz S, Disanto JP, Chelly J, et al. Fine mapping of the human SCIDX1 locus at Xql2-13.1. Hum Mol Genet
1993;2:651-4. 20 Weber JL, Kwitek AE, May PE, et al. Dinucleotide repeat polymorphisms at the DXS453, DXS454 and DXS458 loci. Nucleic Acids Res 1990,18:4037. 21 Roustan P, Curtis ARJ, Kamakari S, et al. Dinucleotide repeat polymorphism at the DXS559 locus Hum Mol Genet 1992;1:778. 22 Graeber MB, Monaco AP, Chelly J, Muller U. Isolation of DNTR repeats from yeast artificial chromosomes encompassing X chromosomal loci PGK1 and DXS56. Hum Genet 1992;90:270-4. 23 Ram KT, Barker DF, Puck JM. Dinucleotide repeat polymorphism at the DXYS1X locus. Nucleic Acids Res 1992,20: 1428. 24 Browne DL, Zonana J, Litt M. Dinucleotide repeat polymorphism at the DXS441 locus. Nucleic Acids Res 1991;19:1721. 25 4th X Chromosome Workshop Report 1993 (in press). 26 Lafreniere RG, Brown CJ, Powers VE, et al. Physical mapping of 60 DNA markers in the p21.1 -q21.3 region of the human X chromosome. Genomics 1991;11:352-63. 27 Corcos IA, Lafreniere RG, Begy CR, et al. Refined localization of human Connexin 32 gene locus, GJB1, to Xql3.1. Genomics 1992;13:479-80. 28 Brown CJ, Sekiguchi T, Nishimoto I, Willard HF. Regional localization of CCG1 gene which complements hamster cell cycle mutation BN462 to Xql 1 -Xq 13. Somat Cell Mol Genet 1989;15:93-6. 29 Sekiguchi T, Yoshida MC, Sekiguchi M, Nishimoto I. Isolation of a human X chromosome-linked gene essential for progression from GI to S phase of the cell cycle. Exp Cell Res 1987;169:395-407. 30 Dermietzel R, Spray DC. Gap junctions in the brain: where, what type, how many and why? Trends Genet 1993;16: 186-92. 31 Bergoffen J, Scherer SS, Wang S, et al. Connexin mutations in X linked Charcot-Marie-Tooth disease. Science 1 993;262 :2039-42.
